Life Science Journal 2015;12(4s)
http://www.lifesciencesite.com
Quality of Guava Whey Beverage Fortified With Moringa Oleifera Leaves Extract
Afaf O. Ali, Wael H. M. Elreffaei, Adel M. Elkarmany and Fatma M. A. Elsheek
Regional Center for Food & Feed, Agriculture Research Center, Giza, Egypt.
[email protected]
Abstract: Efficacy of preservation and antioxidant activities of aqueous leaf extract of Moringa oleifera in whey
beverage was determined. It is well known for its nutritional and health benefits and is being recommended for
malnourished people all over the world. Moringa oleifera leaves, is a rich source of bioactive compounds, as
polyphenols. The present investigation aimed to evaluate the stability of bioactive compounds and free radical
scavenging abilities of the leaf extracts guava in whey beverages stored under different conditions. Moringa leaves
were turned into an aqueous extract and fortified to whey guava beverage at different ratios (2.5,5.0, and 7.5%). The
total phenolic and flavonoid contents have been measured using colorimetric methods. The antioxidant capacities
were evaluated using scavenging assays of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) in aqueous extract. The
results showed that extractability of Moringa leaves compounds Total Phenolic Compound (TPP) was significantly
higher (p ≤ 0.05) in the phenolic extract in the beverage contains 7.5% (5.560 ±0. 21g/100ml) of Moringa leaves
extract at them in control without Moringa extract (2.083±0.29 g/100ml) and it showed higher antioxidant activities
at zero time and up to 10 storage days. Consistent differences (P≤ 0.05) in DPPH activity were observed among the
different ratio of Moringa leaves extract in the beverages about 77 to 99% scavenging activity against free radicals
from DPPH at the zero time. Generally, antioxidative potential was expressed at a later stage resulting in higher
DPPH scavenging activity values initially (zero time) and a latter (10 days) decrease in the most of prepared
beverages except for fortified Moringa leaves extract with guava whey at concentration 2.5%, 5.0% and 7.5%. The
sensory evaluation of the whey guava beverage fortified with 2.5 or 5% was the acceptable ratios for using Moringa
leaves extract in such beverage. It is can concluded that, of the stability of the leaf extract of M. oleifera as a
preservative has been limited up to 10 days when uses in the beverages.
[Afaf O. Ali, Wael H. M. Elreffaei, Adel M. Elkarmany and Fatma M. A. Elsheek. Quality of Guava Whey
Beverage Fortified With Moringa Oleifera Leaves Extract. Life Sci J 2015;12(4s):113-122]. (ISSN:1097-8135).
http://www.lifesciencesite.com. 1
Key words: Moringa oleifera, leaves,total phenolic, DPPH, whey beverages
free radicals that may cause some chronic diseases,
including cancer, cardiovascular diseases and cataract
(Li et al., 2001).
Moringa oleifera Lam, commonly referred to as
‘drumstick tree’ (describing of its pods) or horseradish
tree (describing the taste of its roots) also it named
such as Benz olive, Marango and miracle tree, is a
member of Moringaceae family. It was utilized by the
ancient Roman, Greeks and Egyptians (Fahey, 2005).
Moringa oleifera also have an economical importance
in several industrial and medical uses (Nautiyal and
Venhatarman, 1987). Moringa oleifera Lam. is
commonly known in Egypt. Most of the parts of the
plant possess antimicrobial activity (Caceres et al.,
1991).
Almost all parts of this plant have been used
such as seeds, leaves and flowers having as antibiotic,
antitrypanosomal,
hypotensive,
antispasmodic,
antiulcer, anti-inflammatory, hypocholesterolemic,
hypoglycemic activities (Sreelatha et al., 2011) and
cardiovascular and liver diseases, immune boosting
agent, regulate blood sugar and cholesterol (Limaye et
al., 1995). Jung (2014) has shown that an aqueous
extract of M. oleifera leaves exhibited significant
1. Introduction
The uses of plant and herb extracts as
antimicrobial agents in food and soft drinks have also
been reported for centuries (Gulmez et al., 2006). Due
to potential toxicity of chemical food preservatives,
there has been increased demand for food
preservatives from natural sources. This has led
researchers and food processors to come across
natural food additives with a wide range of
antimicrobial activities. As a result, today plant
antimicrobial products have acquired importance in
the food system to retard bacterial and fungal growth
(Souza et al., 2005b). Naturally occurring
antimicrobial compounds isolated from animal, plant
and microbial sources can be used alone or in
combination with other approved antimicrobial
preservatives (Davidson, 2006). In recent years, it has
been shown that a large number of naturally occurring
compounds from vegetables and fruits are effective as
chemo preventive agents. An advantage of diet –
derived products for cancer prevention is that they
also have apparent benefit in other chronic diseases
(Sporn and Suh, 2000 and Kellof, 2000). Therefore,
natural antioxidants can protect the human body from
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antineoplastic activity against a lung cancer cell line
and several other types of cancer cells. The extract
induced apoptosis, inhibited tumor cell growth, and
lowered the internal level of reactive oxygen species
in human lung cancer cells.
They are well known for their pharmacological
actions too and are used for the traditional treatment
of diabetes mellitus (Babu and Chaudhuri, 2005)
hepatotoxicity (Ruckmani et al., 1998), rheumatism,
and venomous bites and for cardiac stimulation
(Chaudhary and Chopra, 1996). WHO has emphasized
strongly on the rational use of traditional and natural
indigenous medicines, for treating diabetes mellitus
(WHO, 1994). The aqueous extract of the leaves has
been found to possess antifertility activity (Prakash,
1998) and is very useful in regulating the thyroid
hormone status in adult Swiss rats (Tahiliani and Kar,
2000).
M. oleifera is rich in various phytochemicals like
carotenoids, vitamins, minerals, amino acids, sterols,
glycosides,
alkaloids,
flavonoids,
moringine,
moringinine, phytoestrogens caffeoylquinic acids and
phenolics in flowers, leaves, roots, fruits and seeds
(Anwar et al., 2007 and Vongsak et al., 2014). In
addition, it contains vitamin B and several amino
acids which could be consumption of nutritious
vegetable recommended for children to overcome
malnutrition in some countries (Fuglie, 2001).
Moringa leaves are quite rich in minerals; however,
their bioavailability is likely to be reduced by the
presence of oxalates and phytates at concentrations of
4.1% and 3.1%, respectively (Foidl et al., 2001). The
polyphenols have antioxidant properties as they could
neutralize or quench oxidants (Pietta, 2000).
Several low molecular weight bioactive
compounds from Moringa seeds with bactericidal,
fungicidal and immunosuppressive activities were
identified (Mahajan and Mehta, 2010). Recent
research has noted that synergistic properties between
individual bioactive compounds in M. oleifera leaves
act in broad aspects of physiology, such as nutrient
absorption and processing, and antioxidant action that
have potential therapeutic effects (Mbikay, 2012).
Whey is typically drain after the cheese curd
forms, being what remains of the milk once the cheese
or casein is removed. Generally 100 L of milk is
produced 87 L of whey is made as a by-product.
Whey contains a nutritional component with
biological value, such as α-LA, β-LG, and lactoferrin
that help reduce health risks (Smithers et al., 1996).
Cheese whey also contains appreciable quantities of
lactic and citric acids, non-protein nitrogen
compounds (like urea and uric acid) and B group
vitamins (Panesar et al., 2007). Whey is also the
highest natural source of branched-chain amino
acids—isoleucine, leucine, and valine and a major
source of glutamine (Joseph et al., 2012). Both
branched-chain amino acids and glutamine play a vital
role in proper cell function, muscle growth, and
protein utilization. Glutamine also feeds the white
blood cells throughout the body and the cells that line
the intestines (a key center of immune health sweet
whey with a pH value between 6 and 7 (Bylund,
1995). Moreover, whey proteins have been used as
ingredients in infant formulas, sport bars, bakery
products, and yoghurt. Whey has been incorporated
into fruit drinks, which were favorably received by
consumers (Shukla et al., 2004). The protein ratio is
about 0.65, lactose 5.2 and 93% moisture.
Therefore, the aim of this study was to (i)
investigate the effects of addition different levels of
Moringaoleifera leaves aqueous extract on the
phenolic
compounds,
antioxidant
activity,
preservative activity and minerals content of guava
whey beverage, and (ii) evaluate physical and sensory
properties of guava whey beverage fortified with
Moringa oleifera leaves aqueous extract.
2. Material and Methods
Fresh sweet whey (7.4% TS and 0.8% protein
0.8%) was obtained from Faculty of Agric., Cairo
Univ., Giza, Egypt. High quality guava fruits were
obtained from local market, Giza, Egypt. Fresh
Moringa leaves were kindly obtained from National
Research Center, Agronomy Dep., Giza, Egypt.
Methods
Moringa extract:
Fresh Moringa leaves were washed in distilled
water to remove the dusts, dried in a hot air-blowing
oven at 50°C and ground to a fine powder in a
mechanical blender. The powders (10 g) were mixed
individually with 100 mL distilled water and
maintained at 100°C using a water bath with constant
stirring (300 rpm) for 2 hr as suggested by Da Silva et
al., (2013). The mixtures were cooled to room
temperature, stored overnight at 4.0±2°C and then
filtered through 40-mesh screen. The filtrate was
concentrated to 50.0% of initial volume at 50°C using
a rotary vacuum evaporator (ROTAVAPOR R110,
Buchi, Switzerland) and stored at -20°C until used.
Guava juice:
Fresh high quality guava fruits was well washed,
cut to pieces and then pulped in a blender. The
homogenized mixtures of was filtered through a
muslin cloth, the resultant juice was filled into
polyethylene bags and stored at -18°C until used.
Preparation of whey beverages:
Five percent of sugar and 0.2g of carboxey
methyl cellulose (CMC) were added to sweet whey.
The mixture were heated to80°C, cooled to 37°C.
Then 20% (w/w) of guava juice were added. 2.5, 5.0
and 7.5% Moringa extract were added separately to
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serve 3 treatment A, B, C, two control samples were
conducted :control 1 and control 2. Control 1 was
whey beverage with guava juice without moringa
extract. Control 2 was whey beverage with 2.5%
Moringa extract without guava juice. All sample were
stored at 4± 1°C for 10 days. All treatment were
conducted in three replicates
Chemical composition analysis:
Whey beverage was dried at 60 °C up to weight
stable to obtained solid content. Moisture, total
nitrogen, fat, ash and fiber contents of whey beverage
samples were determined according the method of
AOAC (2012). The protein content was obtained by
multiplying the percentage of TN by 6.25. Available
carbohydrate content was obtained by difference in
order to achieve 100 g/100 g of total composition
(FAO, 2003). The pH value was measured using
digital pH meter (HANNA, Instrument, Portugal) with
glass electrode.
Mineral content of the minerals Mn, K, Cu, Fe
and Mg were then run through an Atomic Absorption
Spectrophotometer (Varian, AA240, Victoria,
Australia) using air acetylene flame to determine the
mineral content according to AOAC (2012).
Methanol extracts:
In order to separate the phenols in aqueous
extracts, the method of Ramey et al., (1986) was used.
All dried samples of beverage were milled, 50 g of
each dried sample solved in 200 ml of methanol and
shaked for continuous 3 days. These extracts were
subjected to filtrate and evaporated by rotary at 40-50
°C. Obtained dried phenols were redissolved in
methanol, after evaporation of the solvent, for
subsequent chromatographic analysis.
GC–MS profile:
The GC–MS instrument used to separate and
detect methanol-extracting beverages according to the
method described by AOAC (2012) and Boskou
(2005). One gram of dried sample was extracted three
times with methanol l2 ml. The extracted was
combined and methanol evaporated under reduced
pressure. The residue was dissolved in acetonitrile
(2ml) and washed two times with hexane (3ml).
Acetonitrile was evaporated under vacuum and the
residue was dissolved in methanol (1ml). Injections of
10µl from this dissolve extracted lipid in methanol,
were performed into using a GC/MS (Agilent
Technologies 6890N computerized system coupled to
an MSD, Agilent 5973B mass spectrometer).
Total phenolic content:
The total phenolic content of the Moringa extract
and sample of guava juice with and without Moringa
extract at different storage periods were determined
according to Folin Ciocalteu's method (Swain and
Hillis, 1959) with some modifications. In a vial, 50 μL
of extract, 800 μL distilled water and 25 μL (0.25 N)
Folin Ciocalteu's reagent were mixed and incubated at
room temperature for 3 min. Then, 100 μL sodium
carbonate solution (1 N) was added and further
incubated for 2 hr at room temperature. The
absorbance was read at 725 nm using
spectrophotometer (Analytikjena, Win Aspect plus,
Germany). Gallic acid was used in a standard curve
and the results were expressed in terms of Gallic acid
equivalent (GAE g/100ml).
Antioxidant activity by DPPH (2, 2-Diphenyl-1Picrylhydrazyl):
The Antioxidant activity of the extract was
measured in terms of hydrogen donating or radical
scavenging ability using the stable free radical DPPH
(Pothitirat et al., 2009). One-milliliter solution of the
extract in methanol was added to 0.5 ml of 0.15 mM
DPPH solution in methanol. The contents were mixed
vigorously and allowed to stand at 20°C for 30 min.
The absorbance was measured at 517 nm using
spectrophotometer (Analytikjena, Win Aspect plus,
Germany). Scavenging activity (SA) of the DPPH free
radical was determined from the curve of percent
scavenging plotted against the concentration. The
capability to scavenge the DPPH radical was
calculated using the following equation:
DPPH scavenging effect % = [(A0 - A1/A0) - 100],
Where: A0, was the absorbance of the control reaction
and A1, the absorbance in the presence of the sample.
Physical properties:
Viscosity of the whey beverage samples was
determined using a Brookfield digital Rheometer
(Model Melvern, Kinexus, Germany). Readings were
taken at the shear rate 10 s-1 using spindle -40 at
200°C. Apparent viscosity measurements were carried
out at 20 o C ± 0.1 and expressed as x 10 -5m2/s.
Sensory evaluation:
Sensory evaluation was carried out on the guava
and Moringa beverage, using 20 judges selected from
their consistency in scoring and the samples were
evaluated for flavor, color and appearance using grade
of degree 40 (Larmond, 1985).
Statistical analysis:
Analysis of variance (ANOVA) and Duncan's
test were conducted using a Statistical Analyses
System (SAS, 2004). A probability to P ≤ 0.05 was
used to establish the statistical significance.
3. Results and Discussions
Chemical composition:
The mean values of chemical composition of
guava whey beverage fortified with and without
Moringa leaves aqueous extract are shown in Table 1.
The statistical analysis of the five whey beverage
either fortified with guava or Moringa showed that,
there was significant difference at 5% in all beverages
according to type of fortified ratio of Moringa leaves
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extract even though, there were only slight variations
among samples, statistical differences were observed
(P≤ 0.05). A significant difference was observed (P≤
0.05) in fiber, moisture, protein, fat and ash, which
might be expected for this kind of whey beverages,
due to some variations during preparation of whey
beverage. Fiber and protein content were increased in
whey beverage C and D (major contribution to
chemical composition may be attributed to the
Moringa leaves aqueous derived ingredients in these
beverage formulations.
Table (1): Chemical composition of fresh guava whey beverage fortified with and without Moringa leaves aqueous extract.
Guava whey beverages
Chemical composition
Control 1**
Control 2*** A
B
C
Probability (p≤0.05)
6.84±0.11 b
8.09±0.24 a
5.91±0.08 c
5.36±0.02 d 5.84±0.11 c
0.001
Fiber (%)
10.10±0.71 ab 10.57±0.21 a
10.65±0.30 a 9.54±0.34 b 10.80±0.28 a 0.021
Moisture (%)
5.95±0.21b
7.15±0.21 a
5.95±0.35 b
6.25±0.35 b 6.85±0.07 a
0.000
Protein (%)
d
e
c
2.26±0.00
2.03±0.07
3.26±0.11
3.81±0.13 b 4.28±0.11 a
0.000
Fat (%)
1.88±0.08 c
1.86±0.11 c
4.05±0.49 a
4.20±0.18 a 2.72±0.36 b
0.000
Ash (%)
72.97
70.3
70.18
70.84
69.51
Carbohydrates
Mean (n=3± SD) with different letters in the same row imply significant differences at p≤ 0.05; Control 1**: control of guava whey without
Moringa extract, Control 2***: control whey beverage with 2.5% moringa extract and without guava juice; A, guava whey beverage fortified
with 2.5% Moringa extract; B, guava whey beverage fortified with 5.0% Moringa extract; C, guava whey beverage fortified with 7.5% Moringa
extract.
beverage without Moringa without guava (D) had a
lower than control beverage in potassium content.
Regards results, Moringa extract is contains a higher
ratio of macro nutrient elements, which remain in
their fortified whey beverages. These results also
indicated by Gowrishankar et al., (2010) that the
therapeutic effects of Moringa leaves have been
attributed to the combined actions of various
bioactive components found in the plant that include
trace metal ions, especially K, Ca, P, Zn, Mg and Fe.
Minerals content:
Table 2 shows the minerals content in guava
whey beverages fortified with and without Moringa
leaves aqueous extract. A positive relation between
addition of aqueous Moringa extract to guava whey
beverage and their mineral content was noted. Whey
beverage containing 2.5% Moringa extract without
guava juice was higher in Mn, Cu and Fe when
compared to the control guava whey beverage
without Moringa extract. Meanwhile, fortified whey
Table (2): Minerals content in fresh guava whey beverage fortified with and without Moringa leaves aqueous extract
Nutrients elements
Guava whey beverage
Mn
K
Cu
Fe
Mg
---------------------------------------- (mg/kg) -----------------------------------------Control 1**
15.49
8554
11.10
53.05
813
Control 2***
88.31
7661
12.63
113.5
1052
(A)
17.49
9739
7.5
65.13
1252
(B)
61.68
8715
9.42
93.35
1004
(C)
74.91
9587
9.52
108.7
1490
Control 1**: control of guava whey without Moringa extract, Contol 2***: control whey beverage with 2.5% moringa extract and without guava
juice; A, guava whey beverage fortified with 2.5% Moringa extract; B, guava whey beverage fortified with 5.0% Moringa extract; C, guava
whey beverage fortified with 7.5% Moringa extract
containing guava and milk whey. Most of these acids
were responsible to decrease pH of control beverage.
Other compound less than 1% are responsible to
flavor of beverage were identified in control beverage
such as lactic acid ethyl ester; nonanoic acid ethyl
ester; and propionic acid, 3 nitro. Also, phenol, 4methyl-2-[5-2-thienyl) pyrazol-3-yl] (3.55%) can
identified in the control beverage as phenolic
antioxidant material. Consider to fortified the control
beverage with 2.5% Moringa extract (control 2)
phenol, 4-methyl-2-[5-2-thienyl) pyrazol-3-yl] was
defiantly increased up to 20.25%. This increased in
phenolic compound in beverage A may attributed to
the content of phenolic compounds in Moringa
extract. Vitamin D2 and Vitamin A were identified in
Phenolic compounds profile:
Moringa leaves are used as herbal medicines and
food supplements because of their natural antioxidant
compounds such as phenolic, flavonoids and vitamin
C (Sahakitpichan et al., 2011). The results of phenolic
compound in the controls and beverage samples are
reported in Table 3.
Considering both control beverage were contains
18-nanadecenoic acid (27.99%), p-menthane-1, 2, 3triol (12.55%) and octadecanoic acid, 4-methyl
(11.55%) as the most three abundant compounds
(table 3). The lignoceric acid, pentadecanedioic acid,
6-octadecenoic acid, 6-Octadecenoic acid and LAscorbic acid, 6-stearate were the second abundant
GC-MS identified compound in control beverage
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equal ratio (4.52%) in the 2.5% Moringa beverage
(A). Both of Butyric acid, 2-hydroxy-3-mthyl, methyl
ester (17.47%) and Propanoic acid, 3-nitro (2.53%)
were identified in beverage (A) which consider as
acids of flavoring material and preserve the product.
Another types of acids could identified in 5.0%
Moringa beverage with guava, acetic acid and
propionic acids were the most abundant acids in
beverage (B) raged 7 up to about 21%.The major
monoterpene hydrocarbons of volatile oil in amounts
less than 3% were Limonene in beverage B.
Table 3 shows about 10 major volatile
compounds as identified in the beverage C. These
compounds include different chemical classes, such as
alcohols, ketones, aldehydes and one heterocyclic
compound. In particular, the identified volatile
compounds were acetic acid, 10 aroma volatiles.
Among these identified compounds2-hexyldecanoic
acid, Dodecanoic acid and Docanoic acid, 2-hexyl
were the most abundant volatile acids in the beverage
C, this is attribute to the increase ratio of aqueous
Moringa extract in whey guava beverage.
Corresponding to add Moringa oleifera extract in
beverage, type of natural volatile antioxidants was
obvious in GC-MS profile. Most of these volatile is
concern
in
2-hexyldecanoic
acid,
2octadecoxyethanol,
p-Methane-1,
2-diol,
and
Geranylisovalerate. These compounds were similar to
identify by Mukunzi, et al., (2011) in Moringa leaves
and Leon-Rodriguez et al., (2008). Control 2 sample,
it is the interested of propionic acid, 3 nitro (41.02%);
hexanoic acid (19.84%), and –ethyl cyclobutanol
(14.70%) were contained as most predominate
identify compounds (Table 3). A wide variety of
essential oils are known to possess antimicrobial
properties and in many cases this activity is due to the
presence of active constituents, mainly attributable to
isoprenes such as monoterpenes, sesquiterpenes and
related alcohols, other hydrocarbons and phenols
(Berger, 2007). In this study, the Moringa oleifera
extract contains hydrocarbon and alcoholic as well as
phenolic, might be consider as microbial inhibition
and increased preservative activity of fortified
beverage by Moringa. The compounds also are
increase self-life of beverage contains Moringa leaves
extract. In addition, it can be deduced that there is a
relationship between the chemical components of the
essential oils and the antimicrobial activity of aqueous
Moringa leaves.
Total phenol content (TP)
Table 4 considerable antioxidant capacity
presents the analytical data for total phenolic of the
studied samples of fortified guava whey with Moringa
extract beverages. The total phenolic compounds of
the extracts were expressed as Gallic acid Equivalent
in g/100ml. Considerable antioxidant capacity was
also detected in five beverages along storage period 5
and 10 days. The fortified beverage with 7.5%
Moringa leaves extract, significantly (P ≤ 0.05)
affected the amount of TP in the (C) (5.56 g/100ml) at
zero time of prepare of beverage. Meanwhile, there is
non- significantly affected between control1 and 2 at
the zero time. At the storage period more than 5 days
the beverage C has a considerable amount of TP and
significantly higher than control1 beverage.
Whereas, TP was significantly (P≤ 0.05)
decreased after 10 days of storage in beverages C and
control 2. However, the beverage C, that was
contained 5.0% Moringa leaves extract significantly
stable and consider as higher in TP at different time of
storage period and other beverages samples. The
obtained results are very consistent with many
previously reported results indicating that phenolic
compounds are generally less soluble in water (Wanga
et al., 2009). It could be concluded that, the total
phenolic contents indicate that most of the potent
antioxidant and phenolic compounds in the beverage
C.
Antioxidant activity:
Consistent differences (P ≤ 0.05) in DPPH
activity were observed among the different ratio of
Moringa leaves extract in the whey beverages (Table
5). About 77 to 99% scavenging activity against free
radicals from DPPH in the zero time were observed.
The radical-scavenging capacities of Moringa
aqueous extracts have been previously observed in
different model systems (Razis et al., 2014 and
Ndhlala et al., 2014).
The Moringa leaves extracts was show a higher
scavenging activity in the fortified beverages with 5.0
and 7.5% Moringa in guava whey beverages
especially a long of storage period (10 days).
Meanwhile, the 2.5% Moringa beverage extract was
quite similar to control in zero and after 5 days of
storage. The positive anti-oxidation and preservation
of Moringa in beverage B and C, its might attributed
to the synergistic role of natural compound and
vitamins were found in these beverages. The
scavenging potential of fortified Moringa leaves
extract was twice greater in B and C than original
beverage without Moringa extract, followed by in
beverage A. Furthermore, the presence of a negligible
concentration of ascorbic and acetic acids in the
respective extracts contributed to the effectiveness of
phenolic-induced reducing power. Moreover, the
hydrophilic antioxidant and ascorbic acid were less
effective in an oil-in-water emulsion system, whereas
the opposite trend was found for the hydrophobic
antioxidants (phenolic acids, flavanones, flavonols,
etc.).
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Table (3): Comparison of phenolic compound in guava whey beverages fortified with and without Moringa
leaves aqueous extract
Phenolic compounds
Propionic acid,3 nitro
Acetic acid, 2-benzolthio. 2-oxo-2-phenylester
Lactic acid, ethyl ester
Cyclohexene, 4-isopropenyl-1-methoxymethoxymethyl
Β-Asarone
Nonanoic acid, ethyl ester
Ocimene
Heptadecanoic acid
Aracdeic acid
Fumaric acid, 2,2-dichloroethyl pentadecyl ester
Eurcic acid
9-Hexadecenoic acid
Phenol, 4—methyl-2-[5-2—thienyl) pyrazol-3-yl]
6-Octadecenoic acid
L-Ascorbic acid, 6-stearate
Pentadecanedioic acid
Lignoceric acid
Octadecanoic acid, 4-methyl
p-Menthane-1,2,3-triol
18-Nanadecenoic acid
Butyric acid, 2-hydroxy-3-mthyl, methyl ester
Acetic acid, ethoxy, ethylester
Propioic acid, 2-hydroxy, ethyl ester
Propanoic acid, 3-nitro
Limonene
Vitamin A aldehyde
-himachaleneα
Patchoulene
Vitamine D2
2,4-Di-tert-butylphenyl benzoate
Propanoic acid, 2,3-dihydroxy-2-methyl
Acetic acid, 2,2-[oxybis (2,1-ethanediyloxy)]bid
Propionic acid
Pentanoic acid
Acetic acid, 2,2-[oxybis (ethanediyloxy)]bis
2-Propenoic acid, tetradecyl ester
p-Methane-1,2-diol
Undecanoic acid
Methol, 1-(butyn-3-one-1-yl),(1R,2S,5R)
Dodecanoic acid
2-octadecoxyethanol
14-pentadecenoic acid
Geranylisovalerate
o-Anisic acid, 3.6 –dichloro-5hydroxy
9-hexadecenic acid
1-hexadecsnol,2-methyl
z-11-tetradecenoic acid
2-methyl-E,E-3-13-Octadecadien-1-ol
Pentadecanoic acid
2-hexyldecanoic acid
n-hexadecanoic acid
Docanoic acid, 2-hexyl
Liolenin, 1-mono
-2Ethyl cyclobutano
)Z)-3-Octenol
3-Methylbutanoic acid
Hexanoic acid
4-Hexenoic acid
E-beta-Ionone
2-Hexenoic acid
Phenyl acetonitrile
Octanoic acid
beta-Ionone epoxide
Control 1**
Area %
RT
0.97
4.6
3.47
4.73
0.89
5.75
0.85
6.69
0.69
8.31
0.77
8.52
0.77
9.08
1.74
10.21
0.97
11.58
2.51
12.23
2.12
12.65
1.74
13.66
3.55
15.53
6.76
16.16
4.63
17.37
7.72
17.53
7.72
18.08
11.58
18.15
12.55
18.93
27.99
19.57
A
Area %
RT
2.53
11.6
24.05
5.06
20.25
12.65
13.66
15.3
17.47
6.33
3.80
2.53
1.27
4.56
3.80
3.80
4.53
4.08
4.15
4.38
4.79
6.75
8.24
10.70
10.87
11.41
B
Area %
6.56
3.28
1.09
2.84
0.00
1.09
3.94
0.00
5.03
18.60
12.04
20.79
17.51
7.22
RT
4.15
4.38
4.79
6.75
C
Area %
RT
1.59
4.79
1.88
3.10
6.38
5.91
5.44
7.32
2.81
5.16
3.75
1.41
4.69
5.63
5.16
5.16
1.88
9.29
3.28
7.97
7.04
5.32
12.76
13.78
15.14
15.45
16.39
16.64
16.80
17.04
17.21
17.46
17.70
17.81
17.98
18.07
18.64
19.27
19.55
19.92
Control 2***
Area %
RT
41.02
4.6
9.66
4.73
10.70
10.87
3.62
4.06
4.26
14.70
15.80
15.9
14.70
0.48
1.61
19.84
0.36
2.20
2.11
0.8
1.15
1.77
8.0
13.9
16.04
17.71
18.38
18.74
18.81
18.61
19.60
19.21
Control 1**: control of guava whey without Moringa extract, Control 2***: control whey beverage with 2.5%
Moringa extract and without guava juice. A: guava whey beverage fortified with 2.5% Moringa extract, B: guava
whey beverage fortified with 5.0% Moringa extract and C: beverage fortified with7.5% Moringa extract.
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Generally, antioxidative potential was
expressed at a later stage resulting in higher DPPH
scavenging activity values initially (zero time) and a
latter (10 days) decrease in the most of prepared
beverages except for fortified Moringa leaves extract
with guava whey at concentration 2.5%, 5.0% and
7.5%. On the other hand Vongsak et al. (2013), the
investigation of chemical stability of the leaf extract of
M. oleifera has been limited.
Table (4): Total phenol content (g/100ml) and relation of progressive stability in guava whey fortified with Moringa extract
Guava whey beverages
Control 1
Control 2
A
B
C
Probability
Total phenol content at different storage periods*
Zero time
5 days
2.083±0.29 C
4.670±0.01 c
2.450±0.06 c
2.363±0.12 d
3.020±0.04 b
4.547±0.15 c
b
3.490±0.11
5.257±0.02 b
5.560±0.21 a
5.747±0.12 a
0.0001
0.0001
10 days
2.717±0.04 c
1.853±0.06 d
4.337±0.08 b
5.353±0.17 a
1.520±0.05 e
0.0001
Mean (n=3± SD) with different letters in the same row imply significant differences at P≤ 0.05; Control 1**: control of guava
whey without Moringa extract, Control 2***: control whey beverage with 2.5% Moringa extract and without guava juice. A:
guava whey beverage fortified with 2.5% Moringa extract, B: guava whey beverage fortified with 5.0% Moringa extract and C:
beverage fortified with 7.5% Moringa extract.
Table (5): Antioxidant activity (scavenging activity SA) in guava whey fortified with and without Moringa leaves aqueous extract
Items
SADPPH• (%)
Storage period
(day)
zero
5
10
Treatments
Cont
79.33±0.35 c
73.48 ±0.91 d
65.93 ±0.48 e
Control 2***
77.02 ±0.07 d
78.50 ±1.04 c
76.41 ±0.57 d
A
76.50 ±0.40 d
85.07 ±1.18 b
86.58 ±1.75 d
B
99.19 ±0.42 a
94.77 ±0.81 a
99.05 ±0.67 a
C
98.27 ±0.34 b
94.09±1.07 a
96.27 ±0.44 b
Probability p≤0.05
0.0001
0.0001
0.0001
Mean (n=3± SD) with different letters in the same row imply significant differences at P≤ 0.05; Control 1**: control of guava
whey without Moringa extract, Control 2***: control whey beverage with 2.5% Moringa extract and without guava juice A:
guava whey beverage fortified with 2.5% Moringa extract, B: guava whey beverage fortified with 5.0% Moringa extract and C:
beverage fortified with7.5% Moringa extract
whey beverages fortified with Moringa extract ranged
from 200 to 240 (x 10 -5m2/s) at fresh time and ranged
from 190 to 500 (x 10 -5m2/s) at 10 day. The average
of viscosity in beverage C (7.5% Moringa with guava
whey beverage) was the highest viscosity values at the
different storage periods. Viscosity also tends to
increase during refrigerated storage, which could be
attributed to the behavior of the beverage during
cooling can discussed based exclusively on the
characteristics of its carboxy methyl cellulose (CMC),
but also on the influence of protein of sweet whey on
the beverage.
The protein in the beverage resulted increase of
viscosity depend on molecular structure of whey
protein in the beverages. This is in agreement by
Oszvald et al. (2007). According to the characteristics
of Moringa leaves extract the forecast of behavior was
an increase in viscosity due to CMC and flocculation
activity of Moringa leaves extracts. These results
could be accompanied with the binding of the protein
particles in an emulsion is the result of three types of
forces, i.e. attractive, repulsive, and steric interactions.
The van der Waals attractive interactions are universal
in all food emulsions and are desirable because they
lead to the formation of a network-like structure to
stabilize emulsion products and increase viscosity
(Logaraj et al., 2008).
pH of bevarages fortified guava whey with
Moringa extract:
Fig 1 shows that a gradual increase in pH value
was observed with increasing the Moringa extract
level, while as storage period increased, the pH value
of guava whey beverages fortified Moringa extract
decreased. These positive correlation between
concentration of used Moringa leaves extract in
beverage and pH values according to period zero, 5
and 10 days of storage beverages, was found R2 =
+0.517,
+0.384
and
+0.457,
respectively.
Considerably a significant positive correlation
between concentration of Moringa in the fortified
beverage and pH during storage periods has been
indicated, that aqueous Moringa extract containing
preservative agents to increase in the first period of
storage and it decreased according to storage period.
The fortified guava why beverage with Moringa
aqueous extract was considered as not stable (Fig 2).
pH variations was regular in the fortified Moringa
beverages during storage. After stored 10 days, the
beverage showed small variations in the values of pH.
Viscosity of fortified guava whey with Moringa
extract:
As shown in Table 6, viscosity value was higher
in guava whey fortified with Moringa leaves extract
that non fortified. In particular, the viscosity of guava
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Figure (1): Correlation between concentrations of fortified guava whey with Moringaleaves extract on PH value.
Table (6): Viscosity of guava whey fortified with and without Moringa leaves aqueous extract
Treatments
Storage period
Items
(day)
Control 1
Control 2***
A
B
B
190
40
240
220
220
zero
Viscosity
170
60
360
180
180
5
-5 2
(x 10 m /s)
190
110
190
250
250
10
Control 1**: control of guava whey without Moringa extract, Control 2***: control whey beverage with 2.5% Moringa extract
and without guava juice A: guava whey beverage fortified with 2.5% Moringa extract, B: guava whey beverage fortified with
5.0% Moringa extract and C: beverage fortified with7.5% Moringa extract
Table (7): Sensory evaluation of guava whey fortified with and without Moringa leaves aqueous extract
Treatments
Storage period
Sensory properties
(days)
Cont
Control 2 ***
A
B
C
36
30
38
35
35
zero
30
30
38
35
35
Flavour (40)
5
30
30
36
35
35
10
36
32
36
36
32
zero
32
30
34
36
30
Colour (40)
5
32
30
32
32
30
10
38
30
38
35
35
zero
Appearance
35
30
35
35
32
5
(40)
35
30
35
34
32
10
Control 1**: control of guava whey without Moringa extract, Control 2***: control whey beverage with 2.5% Moringa extract
and without guava juice A: guava whey beverage fortified with 2.5% Moringa extract, B: guava whey beverage fortified with
5.0% Moringa extract and C: beverage fortified with7.5% Moringa extract.
beverage with 2.5 or 5% was the fit ratios for using
Moringa leaves extract in such beverage.
Sensory evaluation:
Sensory evaluation in guava whey beverages
fortified with and without Moringa leaves aqueous
extract is shown in Table 7. After preparation of the
beverages, no major changes occurred in flavor of all
five beverages except for fortified beverage with 7.5%
Moringa extract that recorded decrease in color and
appearance scores compared to control beverage.
Control 2 beverage has lower in the flavor and
appearance at zero time, 5 and 10 days. Both beverage
B and C had shown a little decrease in flavor, color
and appearance in zero time up to 10 days. Beverages
A and B gained higher scores in flavor, color and
appearance throughout storage period. This result
indicated that, the fortification of whey guava
Conclusion
The Moringa oleifera extract contains
hydrocarbon and alcoholic as well as phenolic, might
be consider as microbial inhibition and increased
preservative activity of fortified beverage by Moringa.
These compounds also, are increase self-life of
beverage containing Moringa leaves extract.
Fortification of whey with guava juice and Moringa
leaves aqueous extract produced beverage with higher
nutrients like protein, fiber and mineral contents.
Also, the guava whey beverage fortified with 5.0%
Moringa extract was more stable and consider as
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higher in TP and antioxidant activity at different time
of storage period.
13.
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